Protocol for generating reproducible miniaturized controlled midbrain organoids
Summary: Here, we present a protocol for generating miniaturized controlled midbrain organoids (MiCOs) of reproducible size and cellular composition, without a necrotic center. We describe steps for maintaining and passaging human pluripotent stem cells, generating MiCOs using AggreWellTM400, and ma...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723004185 |
| _version_ | 1797774509820870656 |
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| author | Muwan Chen Jonathan Christos Niclis Mark Denham |
| author_facet | Muwan Chen Jonathan Christos Niclis Mark Denham |
| author_sort | Muwan Chen |
| collection | DOAJ |
| description | Summary: Here, we present a protocol for generating miniaturized controlled midbrain organoids (MiCOs) of reproducible size and cellular composition, without a necrotic center. We describe steps for maintaining and passaging human pluripotent stem cells, generating MiCOs using AggreWellTM400, and maintaining them in an EB-Disk360on an orbital shaker, eliminating the need for Matrigel or a spinner flask and preventing organoid fusion. We then detail organoid collection for different endpoint analysis. This protocol is suitable for compound screening and disease modeling studies. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-12T22:22:10Z |
| format | Article |
| id | doaj.art-d16395151bc94671b04ab9481ec53b25 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-12T22:22:10Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-d16395151bc94671b04ab9481ec53b252023-07-23T04:56:17ZengElsevierSTAR Protocols2666-16672023-09-0143102451Protocol for generating reproducible miniaturized controlled midbrain organoidsMuwan Chen0Jonathan Christos Niclis1Mark Denham2Danish Research Institute of Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Aarhus University, 8000C Aarhus, Denmark; Department of Biomedicine, Aarhus University, 8000C Aarhus, Denmark; Corresponding authorCell Therapy R&D, Novo Nordisk, 2760 Måløv, DenmarkDanish Research Institute of Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Aarhus University, 8000C Aarhus, Denmark; Department of Biomedicine, Aarhus University, 8000C Aarhus, Denmark; Corresponding authorSummary: Here, we present a protocol for generating miniaturized controlled midbrain organoids (MiCOs) of reproducible size and cellular composition, without a necrotic center. We describe steps for maintaining and passaging human pluripotent stem cells, generating MiCOs using AggreWellTM400, and maintaining them in an EB-Disk360on an orbital shaker, eliminating the need for Matrigel or a spinner flask and preventing organoid fusion. We then detail organoid collection for different endpoint analysis. This protocol is suitable for compound screening and disease modeling studies. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723004185NeuroscienceOrganoids |
| spellingShingle | Muwan Chen Jonathan Christos Niclis Mark Denham Protocol for generating reproducible miniaturized controlled midbrain organoids STAR Protocols Neuroscience Organoids |
| title | Protocol for generating reproducible miniaturized controlled midbrain organoids |
| title_full | Protocol for generating reproducible miniaturized controlled midbrain organoids |
| title_fullStr | Protocol for generating reproducible miniaturized controlled midbrain organoids |
| title_full_unstemmed | Protocol for generating reproducible miniaturized controlled midbrain organoids |
| title_short | Protocol for generating reproducible miniaturized controlled midbrain organoids |
| title_sort | protocol for generating reproducible miniaturized controlled midbrain organoids |
| topic | Neuroscience Organoids |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723004185 |
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